Six Holstein calves, 3 to 4 weeks of age were fed a milk replacer and a starter concentrate. Calves were slaughtered after 105 days, without fasting. Contents of the rumen, abomasum, duodenum, jejunum + ileum, caecum and colon were weighed, the pH was measured and the rate of hydrolysis of emulsified tributyrin determined. In all calves, the highest and the lowest specific lipolytic activity (per g digesta) were found in the duodenum and abomasum, respectively. The ruminal lipolytic activity was rather low, but thank to its great volume the rumen contained almost one half of the total lipolytic activity present in the digestive tract. The reflex of oesophagal groove closure, however, restricts the potential role of the rumen in digestion of dietary fat to hydrolysis of lipids present in the solid feed. In pooled samples of digesta from the rumen, abomasum, duodenum and jejunum/ileum the highest lipolytic activity was observed at pH of 5.5, 2.5, 8.5 and 9.0, respectively. There was no clear pH optimum for hydrolysis of tributyrin by the caecal/colonic contents. It can be concluded that lipolytic enzymes are present in all sections of the calf digestive tract, but individual digestive segments differ substantially in specific and total lipolytic activity, and in nutritional significance of these activities for the animal.